An Alternative Method for Extracting Plasmodium DNA from EDTA Whole Blood for Malaria Diagnosis

• Published in: Korean journal of parasitology 2018-02, Vol.56 (1), p.25-32
• Main Authors: Seesui, Krongkaew, Imtawil, Kanokwan, Chanetmahun, Phimphakon, Laummaunwai, Porntip, Boonmars, Thidarut
• Format: Article
• Language: English
• Subjects: Original
• ISSN: 0023-4001; 1738-0006
• DOI: 10.3347/kjp.2018.56.1.25

Molecular techniques have been introduced for malaria diagnosis because they offer greater sensitivity and specificity than microscopic examinations. Therefore, DNA isolation methods have been developed for easy preparation and cost effectiveness. The present study described a simple protocol for DNA isolation from EDTA-whole blood. This study demonstrated that after heating infected blood samples with Tris-EDTA buffer and proteinase K solution, without isolation and purification steps, the supernatant can be used as a DNA template for amplification by PCR. The sensitivity of the extracted DNA of and was separately analyzed by both PCR and semi-nested PCR (Sn-PCR). The results revealed that for PCR the limit of detection was 40 parasites/μl for and 35.2 parasites/μl for , whereas for Sn-PCR the limit of detection was 1.6 parasites/μl for and 1.4 parasites/μl for . This new method was then verified by DNA extraction of whole blood from 11 asymptomatic Myanmar migrant workers and analyzed by Sn-PCR. The results revealed that DNA can be extracted from all samples, and there were 2 positive samples for ( and ). Therefore, the protocol can be an alternative method for DNA extraction in laboratories with limited resources and a lack of trained technicians for malaria diagnosis. In addition, this protocol can be applied for subclinical cases, and this will be helpful for epidemiology and control.